Stable isotope dilution analysis of the Fusarium mycotoxins deoxynivalenol and 3‐acetyldeoxynivalenol

Abstract

Trichothecenes are secondary metabolites produced by several fungi of the Fusarium genus during their growth period. They inhibit protein biosynthesis in eukaryotic cells resulting in numerous toxic effects such as diarrhea, vomiting, and gastro-intestinal inflammation. Considering its occurrence in food and feedstuff, deoxynivalenol (DON) is one of the most important trichothecenes. We report the synthesis of stable isotope labeled 15-d(1)-deoxynivalenol (15-d(1)-DON) from its natural precursor 3-acetyldeoxynivalenol (3-AcDON) as starting material. Furthermore, a method for the analysis of DON and 3-AcDON using HPLC-MS/MS with stable isotope labeled 15-d(1)-DON and 3-d(3)-AcDON as internal standards has been developed. In total, 18 cereal product samples were analyzed with contamination levels ranging from 10-301 microg/kg for DON and 5-14 microg/kg for 3-AcDON. This is the first report of an isotope dilution MS method for the analysis of type B-trichothecenes.

Determination of deoxynivalenol-sulfonate (DONS) in cereals by hydrophilic interaction chromatography coupled to tandem mass spectrometry

“…Synthesis of 9,15-d 2 -DONS One mg 15-d 1 -DON synthesized in our laboratory (Bretz et al 2006) was dissolved in 1 mL D 2 O and 200 mg SBS were added. After stirring overnight, the reaction mixture was purified using a SepPak C 18 column and 9,15-d 2 -DONS was isolated by preparative HPLC (for parameters, see section "Preparation and purification of DONS").…”

Analysis of Fusarium toxins via HPLC-MS/MS multimethods: matrix effects and strategies for compensation

“…The first is more elaborate as every single sample has to be spiked on several concentration levels. Isotope-labeled IS added to the solid sample prior to cleanup can compensate-in an ideal case-both the losses induced by extraction, cleanup and HPLC analysis as stated by several research teams Rychlik 2006, 2007;Beyer et al 2009;Bretz et al 2006;Haeubl et al 2006Haeubl et al , 2007. However, using isotope-labeled IS for every analyte from the beginning of the sample preparation results in very high costs which are not suitable for routine analysis as well.…”

“…Previously published methods for the determination of trichothecenes, fumonisins and/or zearalenone either did not include the compensation of such effects (Berthiller et al 2005;Cavaliere et al 2005) or included matrix calibration (Biselli et al 2005;Spanjer et al 2008) which is not feasible for routine work. Some other methods describe the use of one internal standard (IS) for several analytes (Berger et al 1999;Kloetzel et al 2006 (European Commission 1999, 2000a, b, 2002 often describe the use of isotope-labeled IS for compensation of matrix suppression or enhancement Rychlik 2006, 2007;Haeubl et al 2006Haeubl et al , 2007Herebian et al 2009;Rychlik and Asam 2008;Bretz et al 2006;Cramer et al 2007), which is in general an effective method, but very expensive as spiking is commonly performed before extraction. Some of these studies reveal that significantly lower values were obtained when no isotope-labeled IS was used (Haeubl et al 2006;Asam and Rychlik 2007), however, there is only less data about the part of matrix effects which is only caused by ionization.…”

Development of a recombinant Fab-fragment based electrochemical immunosensor for deoxynivalenol detection in food samples

“…While different sample treatments were considered, as the regulatory limit for DON is high (1750 g/kg), a simple procedure involving the use of pure water as extraction solvent, which has been previously reported to be extremely efficient for polar trichothecenes such as DON and NIV, was assessed (Bretz et al, 2006;Cahill et al, 1999). For the evaluation of the matrix effects, a water extract obtained by Fig.…”

“…Five different sample treatment procedures (Scheme SI1) were investigated. Extracting with a solution of acetonitrile/water was studied and the possible utilization of a clean-up step, using Solid Phase Extraction (Mycosep or Bondelut cartridges) was considered (Bretz et al, 2006;Milanez and Valente-Soares, 2006). Methanol was also used as an extraction solvent followed by a Bond Elut clean-up procedure.…”